Effective Amounts of (3aR)-1,3a,8-Trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo [2,3-b]indol-5-yl Phenylcarbamate and Methods of Treating or Preventing Neurodegeneration

ABSTRACT

The invention includes an amount of (3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-yl phenylcarbamate for administering to a subject and also a method of preventing or treating neurotoxicity or neurodegenerative processes in a subject in need thereof using the amount thereof.

BACKGROUND OF THE INVENTION Neurotoxic Precipitating Fibrillar Proteins

Neurodegenerative diseases generally affect abstract thinking, skilledmovements, emotional feelings, cognition, memory and other abilities.Despite differences in clinical symptoms and disease progression,disorders from this group share key common features: most of them haveboth sporadic and inherited origins, all of them appear later in life(usually after the fourth or fifth decade of the subject's life), andtheir pathology is characterized by neuronal loss and synapticabnormalities. Until recently, no common molecular mechanism had beenidentified among these diseases. However, various neurodegenerativediseases, such as Alzheimer's disease (AD), Parkinson's disease (PD),Huntington's disease (HD), transmissible spongiform encephalopathies(TSEs), and amyotrophic lateral sclerosis (ALS), have been shown toshare a common cause and pathological mechanism—the misfolding,aggregation and accumulation of proteins in the brain, resulting inneuronal apoptosis. The hallmark feature of conformational disorders isthat a particular protein folds into a stable alternative conformation,which in most cases results in its aggregation and accumulation intissues as fibrillar deposits. These deposits have similarmorphological, structural and staining characteristics (FIGS. 3 and 4).Multidisciplinary studies strongly support this shared cause andpathological mechanism, suggesting that there may be a common therapyfor these devastating disorders.

Mutations in the genes that encode the protein components of fibrillaraggregates are genetically associated with the inherited forms of allneurodegenerative diseases. The familial forms usually have an earlieronset and greater severity than sporadic cases and are also associatedwith a greater amount of protein aggregates (Soto, 2003, Nature Rev.4:49).

Neurotoxic aggregating proteins have not only a common aggregatingpathway, but also common regulatory pathways for their transcription andtranslation. While their transcription is activated by copper and/orzinc ions (Bush et al., 2003, Proc. Natl. Acad. Sci. U.S.A.100(20):11193-94), their translation is upregulated by iron anddown-regulated by iron regulatory protein 1 (IRP1) (FIG. 6A).Specifically, their mRNAs are regulated via the 5′-untranslated region(5′UTR) of their transcript, which folds into a unique RNA stem loopwith a CAGUGN apical loop similar to that encoded in the canonicaliron-responsive element (IRE) of L- & H-ferritin mRNAs (FIGS. 7A, 7B, 9Band 12). IRP1 binds to this IRE stem look and inhibits the translationof the mRNA by the ribosome (Cho et al., 2010, J. Biol. Chem.285(40:31217; FIG. 13). Binding assays have been developed based onthese protein interactions (FIG. 8).

Examples of neurotoxic aggregating proteins are APP (amyloid precursorprotein), Aβ (amyloid-β peptide, a fragment of APP), SOD (super oxidedismutase) proteins, Tau, alpha-synuclein (SNCA), transmissiblespongiform encephalopathy (TSE) prions, and huntingtin (HTT).

In humans, alpha-synuclein is encoded by the SNCA gene (FIG. 11). Analpha-synuclein fragment, known as the non-Aβ component (NAC) ofAlzheimer's disease amyloid and originally found in an amyloid-enrichedfraction, is shown to be a fragment of its precursor protein, NACP (nowreferred to as human alpha-synuclein).

Even though tau is not a member of the iron regulated neurotoxic proteinfamily, it is overexpressed and aggregates in a number of tauopathies.Conditions in which neurofibrillary tangles are commonly observedinclude: Alzheimer's disease, progressive supranuclear palsy, dementiapugilistica (chronic traumatic encephalopathy), frontotemporal dementiaand parkinsonism linked to chromosome 17, Lytico-Bodig disease(Parkinson-dementia complex of Guam), Tangle-predominant dementia, withNFTs similar to AD, but without plaques. Tau deposits tend to appear inthe very old, Picks disease and a number of other neuropathies. Tau'sfibrillary tangles are found in most neurodegenerative disorders.

Transmissible spongiform encephalopathies (TSEs, colectively known asprion diseases) are a group of progressive conditions that affect thebrain and nervous system of mammals, and include devastating diseases asbovine spongiform encephalopathy (BSE, also known as “mad cow disease”)in cattle, and classic Creutzfeldt-Jakob disease, new variantCreutzfeldt-Jakob disease (nvCJD, a human disorder related to mad cowdisease), Gerstmann-Sträussler-Scheinker syndrome, fatal familialinsomnia and kuru (CJD) in humans. Mental and physical abilitiesdeteriorate in the afflicted patients, and myriad tiny holes appear inthe cortex, causing it to appear like a sponge (hence ‘spongiform’) whenbrain tissue obtained at autopsy is examined under a microscope. Thedisorders cause impairment of brain function, including memory changes,personality changes and problems with movement that worsen over time.

Unlike other kinds of infectious disease by microbes, the infectiousagent in TSEs is thought to be a specific protein called prion protein.Misshaped prion proteins carry the disease between individuals and causedeterioration of the brain. TSEs are unique diseases, in that theiretiology may be genetic, sporadic or infectious via ingestion ofinfected foodstuffs and via iatrogenic means (e.g. blood transfusion).Most TSEs are sporadic and occur in an animal with no prion proteinmutation. Inherited TSE occurs in animals carrying a rare mutant prionallele, which expresses prion proteins that contort by themselves intothe disease-causing conformation.

The degenerative tissue damage caused by human prion diseases(Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker syndrome, andkuru) is characterised by four features: spongiform change, neuronalloss, astrocytosis and amyloid plaque formation. These neuropathologicalfeatures have formed the basis of the histological diagnosis of humanprion diseases for many years, although it was recognized that thesechanges are enormously variable both from case to case and within thecentral nervous system in individual cases.

Transmissible spongiform encephalopathies encompass the followingdiseases (including natural host and prion name): scrapie (sheep andgoats; scrapie prion); transmissible mink encephalopathy (TME) (mink;TME prion); chronic wasting disease (CWD) (elk, white-tailed deer, muledeer and red deer; CWD prion); bovine spongiform encephalopathy (BSE)commonly known as “mad cow disease” (cattle; BSE prion); felinespongiform encephalopathy (FSE) (cats; FSE prion); Exotic ungulateencephalopathy (EUE) (nyala and greater kudu; EUE prion); kuru (human;Kuru prion); Creutzfeldt-Jakob disease (CJD) or VariantCreutzfeldt-Jakob disease (vCJD, nvCJD) (human; CJD and vCJD prions);Gerstmann-Sträussler-Scheinker syndrome (GSS) (human; GSS prion); andfatal familial insomnia (FFI) (human; FFI prion).

Alzheimer's Disease

Alzheimer's disease (AD) is the most common progressive dementiaassociated with aging. The cholinergic system is the earliest and mostprofoundly affected neurotransmitter system in AD, with substantiallosses in the forebrain, cortex and hippocampus, which are critical inthe acquisition, processing and storage of memories (Terry et al., 1991,Ann. Neural. 30:572-80; Giacobini, In “Alzheimer's Disease: MolecularBiology to Therapy”; Becker & Giacobini, Eds.; Birkhauser: Boston, 1997;pp 188-204; Becker et al., In “Alzheimer's Disease: from MolecularBiology to Therapy”; Becker & Giacobini, Eds.; Birkhauser: Boston, 1997;pp 257-66).

The major neuropathological hallmarks of AD are β-amyloid plaques,neurofibrillary tangles, and synaptic loss (Khachaturian, 1985, Arch.Neural. 42:1095-1105; FIG. 1). In particular, amyloid-β precursorprotein (APP) is cleaved into a number of toxic peptides, one of thembeing amyloid-β (Aβ): a hydrophobic, neurotoxic self-aggregating 40 to42 amino-acid peptide that accumulates preferentially within senileplaques in the brain. Other peptides are also cleaved from theN-terminus and C-terminus end of APPs. These peptides attack multiplepathways of neuronal cell life, leading to synaptic loss and nerve celldeath. This sequence of events induces neuminflammation and leads tocognitive impairment and neurodegeneration (FIG. 2).

In an original hypothesis for AD treatment, inhibition of theaccumulation of Aβ in the brain could positively affect the course ofAD. Recently, this hypothesis has been expanded by recognizing that APPin the absence of trophic factors is shed from the surface of neuronalcells and processed into an amino terminal fragment (N-APP). Thisfragment binds to DR6 receptors and induces nerve cell death (Nicolaevet al., 2009, Nature 457:981-90). Furthermore, C31 (another factorcleaved from the C-terminus end of APP) has been found to cause nervecell degeneration and death in tissue culture cells and in transgenicmice (Galvan et al., 2006, PNAS 103(18):7130-35). Overexpression of C31has been also shown to lead to neuronal degeneration without Aβ toxicityand plaque deposition. In all three cases (cell death triggered byN-APP, Aβ and/or C31 accumulation), reducing APP synthesis could bebeneficial to the preservation of brain cells, by reducing the formationof neurotoxic plaque through the Aβ pathway and by inhibiting theformation of nerve cell-killing toxic N-terminus and C-terminusfragments.

Current Drugs on Market Symptomatic Compounds

Current AD therapeutic drugs on the market include improving cognitiveprocesses by increasing the levels of acetylcholine. The agents thathave demonstrated the greatest promise in preclinical development arecholinesterase inhibitors and cholinomimetics. The best characterizedcholinesterase is acetylcholinesterase (AChE; EC 3.1.1.7; Soreq et al.,“Human Cholinesterases and Anticholinesterases”; Academic Press: NewYork, 1993). AChE selective inhibitors are used in treatment of AD toamplify the action of acetylcholine (ACh) at remaining cholinergicsynapses within the AD brain. In fact, various AChE inhibitors (such asdonepezil, galantamine and rivastigmine) and a NMDA glutamate receptorblocker (memantine) are the only FDA approved drugs for AD, primarilyproviding symptomatic relief to the patient.

Current Compounds in Development Disease Modifying Compounds

A number of potential Alzheimer's DMDs are in Phase II or Phase IIIclinical trials. Two players (Elan-Wyeth-J&J, and Eli Lilly) lead theway as the frontrunners with a monoclonal and a humanized antibodyagainst. Aβ, respectively, in Phase III clinical trials. Compoundscurrently in Phase I or entering Phase II clinical trials includeRinat/Pfizer's humanized monoclonal antibody, Prana's chelator and QRPharma's Posiphen®. Recently various compounds in Phase II/III failed inthe clinic: Lilly's γ-secretase inhibitor, Medivation/Pfizer's Dimebon®,an antihistamine, TransTech/Pfizer's RAGE®, and CoMentis/Astella'santibody.

Posiphen®

Posiphen® (also known as(3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate; Compound (1) hereafter) is a novel AD drug candidatethat is distinct from the AD drugs currently available as well as fromthe AD drugs currently in development. Compound (1) optimally matchesAChE inhibitory activity with APP/Aβ lowering action. Thisdual-mechanism of action allows for disease modification (bysubstantially lowering brain amyloid levels) and immediate symptomaticimprovement (by inhibiting acetyl cholinesterase).

Compound (1) is a small orally active compound that has high blood brainbarrier permeability (7:1) and lowers amyloid-β precursor protein (APP)levels. In cell cultures, Compound (1) was shown to inhibit APPsynthesis and reduce Aβ formation. The AChE inhibition observed forCompound (1) was found to reside on the N¹-desmethyl metabolite ofCompound (1). Because of this characteristic, Compound (1) has slowonset acetylcholinesterase inhibitory action. Compound (1) and itsmetabolites enter the brain readily, with a 2- to 2.5-time longerhalf-life in brain than in plasma, leading to prolonged efficacy.Compound (1) has been shown to have an excellent toxicology profile anda much better safety profile than traditional AChE inhibitors inanimals.

Posiphen® also inhibits the translation of other neurodegenerativeaggregating proteins such as alpha-synuclein, prions and super oxidedismutase due to its effect on IRP1/IRE in the 5′UTR stem looksstructure of their mRNAs. (FIGS. 5, 6, 9A, and 10).

There is a need in the art to identify novel formulations and dosingregimens that allow the administration of Compound (1) or a salt thereofto effectively treat neurodegenerative diseases, such as but not limitedto AD, in a subject. The present invention fills this need.

BRIEF SUMMARY OF THE INVENTION

The invention includes an amount of(3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3b]indol-5-ylphenylcarbamate (Compound (1)) or a salt thereof, wherein administeringthe amount to a subject results in a peak plasma circulating level ofCompound (1) ranging from about 10 ng/mL to about 160 ng/ml in thesubject.

In one embodiment, the peak plasma circulating level ranges from about80 ng/mL to about 160 ng/ml in the subject. In another embodiment, thepeak plasma circulating level is reached within about 6 hours after theadministering. In yet another embodiment, the peak plasma circulatinglevel is reached within about 3 hours after the administering. In yetanother embodiment, the plasma circulating level of Compound (1) isequal to or greater than about 20 ng/mL for at least 12 hours after theadministering. In yet another embodiment, the plasma circulating levelof Compound (1) is equal to or greater than about 20 ng/mL for at least9 hours after the administering. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (2)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (3)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (4)ranging from about 1% to about 9% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a steady state plasma concentration of Compound(1) of at least about 100 ng/ml in the subject. In yet anotherembodiment, the administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in the subject.In yet another embodiment, the administering results in a steady stateplasma concentration of Compound (3) of at least about 10 ng/ml in thesubject. In yet another embodiment, the administering results in asteady state plasma concentration of Compound (4) of at least about 3ng/ml in the subject. In yet another embodiment, the administeringresults in a brain level of Compound (1) that ranges from about 4 toabout 10 times the plasma level of Compound (1) in the subject. In yetanother embodiment, the brain level of Compound (2) ranges from about15% to about 150% of the brain level of Compound (1) in the subject. Inyet another embodiment, the brain level of Compound (3) ranges fromabout 15% to about 150% of the brain level of Compound (1) in thesubject. In yet another embodiment, the brain level of Compound (4) islower than the brain level of Compound (2) or Compound (3) in thesubject. In yet another embodiment, the subject is a human.

The invention also includes a method of inhibiting production of aneurotoxic aggregating protein in a subject. The method comprisesadministering to the subject a pharmaceutical composition comprising apharmaceutically acceptable carrier and an amount of Compound (1) or asalt thereof, wherein administering the composition to the subjectresults in a peak plasma circulating level of Compound (1) ranging fromabout 10 ng/ml to about 160 mg/ml in the subject, whereby production ofthe neurotoxic aggregating protein in the subject is inhibited.

In one embodiment, the neurotoxic aggregating protein is selected fromthe group consisting of APP, Aβ, SOD, Tau, alpha-synuclein (SNCA), NAC,TSE and HTT. In another embodiment, the administering results in areduction equal to or greater than about 25% in a cerebrospinal fluidmarker selected from the group consisting of sAPP α, sAPP β, Tau andpTau in the subject. In yet another embodiment, the administeringresults in a reduction equal to or greater than about 30% in acerebrospinal fluid marker selected from the group consisting of sAPP α,sAPP β, Tau and pTau in the subject.

The invention also includes a method of treating dementia in a subject.The method comprises administering to the subject a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and anamount of Compound (1) or a salt thereof, wherein administering thecomposition to the subject results in a peak plasma circulating level ofCompound (1) ranging from about 10 ng/mL to about 160 ng/ml in thesubject, whereby the dementia in the subject is treated.

In one embodiment, the dementia is Alzheimer's disease. In anotherembodiment, the dementia is selected from the group consisting ofParkinson's disease, Huntington's disease, Prion's disease, AmyloidLateral Sclerosis and a tauopathy. In yet another embodiment, theadministering results in a reduction equal to or greater than about 25%in a cerebrospinal fluid marker selected from the group consisting ofsAPP α, sAPP β, Tau and pTau in the subject. In yet another embodiment,the administering results in a reduction equal to or greater than about30% in a cerebrospinal fluid marker selected from the group consistingof sAPP α, sAPP β, Tau and pTau in the subject.

In one embodiment, the peak plasma circulating level ranges from about80 ng/mL to about 160 ng/ml in the subject. In another embodiment, thepeak plasma circulating level is reached within about 6 hours after theadministering. In yet another embodiment, the peak plasma circulatinglevel is reached within about 3 hours after the administering. In yetanother embodiment, the plasma circulating level of Compound (1) isequal to or greater than about 20 ng/mL for at least 12 hours after theadministering. In yet another embodiment, the plasma circulating levelof Compound (1) is equal to or greater than about 20 ng/mL for at least9 hours after the administering. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (2)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (3)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (4)ranging from about 1% to about 9% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a steady state plasma concentration of Compound(1) of at least about 100 ng/ml in the subject. In yet anotherembodiment, the administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in the subject.In yet another embodiment, the administering results in a steady stateplasma concentration of Compound (3) of at least about 10 ng/ml in thesubject. In yet another embodiment, the administering results in asteady state plasma concentration of Compound (4) of at least about 3ng/ml in the subject. In yet another embodiment, the administeringresults in a brain level of Compound (1) that ranges from about 4 toabout 10 times the plasma level of Compound (1) in the subject. In yetanother embodiment, the brain level of Compound (2) ranges from about15% to about 150% of the brain level of Compound (1) in the subject. Inyet another embodiment, the brain level of Compound (3) ranges fromabout 15% to about 150% of the brain level of Compound (1) in thesubject. In yet another embodiment, the brain level of Compound (4) islower than the brain level of Compound (2) or Compound (3) in thesubject. In yet another embodiment, the subject is a human.

BRIEF DESCRIPTION OF THE DRAWINGS

For the purpose of illustrating the invention, there are depicted in thedrawings certain embodiments of the invention. However, the invention isnot limited to the precise arrangements and instrumentalities of theembodiments depicted in the drawings.

FIG. 1 is a scheme illustrating the correlation between Alzheimer'sdisease, and (a) senile plaque (amyloid-β peptide), (b) neurofibrillarytangles (hyperphosphorylated tau) and (c) synaptic and neuronaldegeneration.

FIG. 2 is a scheme illustrating the downstream events relating to APPprocessing (adapted from Nicolaev et al., 2009, Nature 457:981-90, andGalvan et al., 2006, PNAS 103(18):7130-35).

FIG. 3 is a series of photographs illustrating the role of cerebralaggregates in neurodegenerative diseases (AD: plaques and tangles; PD:Lewy bodies; HT: Huntington inclusions; TSE: prion amyloid plaque; ALS:superoxide dismutase inclusions)

FIG. 4 is a scheme illustrating protein misfolding and aggregation.

FIG. 5, comprising FIGS. 5A-5D, illustrates the finding that Compound(1) reduces newly synthesized APP without altering APP mRNA or totalprotein synthesis. FIG. 5A: 10 min ³⁵S incorporation into APP, inSH-SY-5Y human neuroblastoma cells. FIG. 5B: newly synthesized totalprotein. FIG. 5C: newly synthesized APP. FIG. 5D: APP mRNA.

FIG. 6, comprising FIGS. 6A-6B, illustrate 5′-UTR of APP mRNA. FIG. 6Ais a scheme illustrating the translational regulation of iron and IRP15′-UTR of APP mRNA. FIG. 6B is a scheme illustrating the mechanism ofaction of Compound (1) on 5′UTR of APP mRNA.

FIG. 7, comprising FIGS. 7A-7B, illustrates 5′ UTR sequence homology ofAPP in mammals. HumanAPP mRNA, SEQ ID NO:1; Mouse APP mRNA, SEQ ID NO:2;Rhesus APP, SEQ ID NO:3; Human L-chain, SEQ ID NO:4; Human H-chain, SEQID NO:5; Consensus, SEQ ID NO:6.

FIG. 8 is a scheme illustrating the IRP/IRE binding assay.

FIG. 9, comprising FIGS. 9A-9B, illustrates the concentration-dependentinhibition of APP and alpha-synuclein translation by Compound (1).H-chain, SEQ ID NO:7; L-chain, SEQ ID NO:8; ASYN, SEQ ID NO:9; APP, SEQID NO:10; APP IRE loop, SEQ ID NO:11.

FIG. 10 is a representation of a gel illustrating that the inhibition ofneural alpha-synuclein expression by Compound (1) is potentiated bycellular iron. Compound (1) decreased alpha-Syn levels dose-dependentlyin dopaminergic SH-SY5Y cells, as was reported for APP: The 5′UTRs ofboth APP & SNCA share 50% homology with the IRE H-ferritin mRNA(Friedlich et al., 2007). Quantitative Western blotting established theefficacy of Posiphen on alpha-Syn expression (IC₅₀<5 μM); afterstandardization for β-actin (Rogers et al. 2010).

FIG. 11 is a scheme illustrating the 5′ UTR sequence homology of SNCA inmammals and birds. H sapiens, SEQ ID NO:12; P. troglodytes, SEQ IDNO:12; M. musculus, SEQ ID NO:13; R. novergicus, SEQ ID NO:14; B.Taurus, SEQ ID NO:15; C. lupus, SEQ ID NO:16; G. gallus, SEQ ID NO:17;Ferritin H-chain, SEQ ID NO:18; Ferritin L-chain, SEQ ID NO:19; APP, SEQID NO:20.

FIG. 12 is a scheme illustrating the 5′ UTR stem loop homology of SNCAin mammals and birds.

FIG. 13 is a scheme illustrating that the prion 5′UTR regions acrossspecies have putative IRP1 binding domains. H. sapiens, SEQ ID NO:21; G.gorilla, SEQ ID NO:22; P. troglodytes, SEQ ID NO:23; M. fascicularis,SEQ ID NO:24; light-chain ferritin, SEQ ID NO:25; heavy-chain ferritin,SEQ ID NO:26; APP, SEQ ID NO:27; B. Taurus, SEQ ID NO:28; O. aries, SEQID NO:29; R. norvegicus, SEQ ID NO:30; M. musculus, SEQ ID NO:31;beta-actin, SEQ ID NO:32.

FIG. 14 is a table illustrating the 5′ UTR sequence homology andPosiphen® inhibition across neurotoxic fibrillar proteins.

FIG. 15 is a series of graphs illustrating the in vivo reduction of APP,Aβ₄₀ and Aβ₄₂ levels in mouse brain by Compound (1). Drug wasadministered once daily×21 day: APP—Western Blot, Aβ—Specific ELISA for1-40 and 1-42 forms.

FIG. 16 is a series of graphs illustrating the effects of Compound (1)in Alzheimer APP23 transgenic mice. APP23 transgenic AD mice weretreated with 25 mg of Compound (1) daily for 14 days. Compound (1)lowered APP levels in the brain of transgenic APP23 mice withoutlowering mRNA levels, improved stem cell survival in brain, increasedstem cell migration to the hippocampus and cortex, and supported stemcell differentiation into neurons rather than into astrocytes and glia.

FIG. 17 is a series of graphs illustrating effects of Compound (1) inTs65Dn mouse model of Down Syndrome. Compound (1) lowered APP levels by30% in brain of trisomy Down Syndrome mice (to control—non-DS brainlevels), and achieved a 10:1 ratio of brain to plasma levels. NGF levelswere elevated in brain, suggesting a potential therapeutic use. BrainAChE levels were significantly lowered (50% at Compound (1) 50 mg/kg),suggesting that administration of Compound (1) provides anticholinergicactivity.

FIG. 18 illustrates selected metabolites of Posiphen® [Compound (1) or(3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate]:

-   N¹-nor-Posiphen [Compound (2) or    (3aR)-3a,8-dimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-yl    phenylcarbamate],-   N⁸-nor-Posiphen [Compound (3) or    (3aR)-1,3a-dimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-h]indol-5-yl    phenylcarbamate], or-   N¹,N⁸-nor-Posiphen [Compound (4) or    (3aR)-3a-methyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-yl    phenylcarbamate].

FIG. 19 is a table illustrating the effects of Compound (1) and selectedmetabolites on molecular markers. Compound (1) is metabolized in theliver into three major compounds: Compounds (2), (3) and (4).

FIG. 20, comprising FIGS. 20A-20B, is a series of graphs illustratinginhibition of Aβ₄₂ levels by Compound (1), as a function of drug plasmalevels. FIG. 20A illustrates brain Aβ₁₋₄₂ levels (±SEM) following 21days of oral Compound (1), as a function of dose of Compound (1). FIG.20B illustrates human and mouse plasma levels of Compound (1) after oraladministration.

FIG. 21, comprising FIGS. 21A-21B, is a series of graphs illustratingthe PK of Compound (1) and selected metabolites. FIG. 21A is a graphillustrating rat levels of Compound (1) and selected metabolites inplasma, brain and CSF (cerebrospinal fluid) at steady state. FIG. 21B isa graph illustrating human levels of Compound (1) and selectedmetabolites inn plasma, brain and CSF at steady state.

FIG. 22, comprising FIGS. 22A-22B, is a series of graphs illustratingthe level of Compound (1) and selected metabolites in human plasma (FIG.22A) and CSF (FIG. 22B) as a function of time after administration.

FIG. 23 is a table illustrating levels of biomarkers in CSF fluid forsubjects after 10 days on Compound (1).

FIG. 24 is a graph illustrating the comparison of Tau before and aftertreatment with Compound (1) in healthy volunteers. Compound (1) wasgiven for 10 days to mild cognitive impairment (MCI) patients, lowerstheir Tau levels back to the levels found in healthy volunteers.

FIG. 25, comprising FIGS. 24A and 25B, illustrate the results of thedetermination of plasma metabolites of phenserine in men (FIG. 25A) andof phenserine in women (FIG. 25B), following 15 mg dose of phenserine onday 35 of study. PT: phenserine; N⁸: N⁸-desamine phenserine; N¹:N¹-desmethyl phenserine; N¹,N⁸: N¹,N⁸-desmethyl phenserine. Note thatthe metabolic profile of phenserine and Posiphen are totally different,with phenserine having a half life of less than one hour and theN¹,N⁸-bisnorphenserine resulting in being the active compound. Posiphenhas a half-life in plasma of 5 hours, with Posiphen being the activedrug component and the N¹,N⁸-bisnormetabolite being a minor andnegligible contributor to the activity.

FIG. 26 is a series of tables summarizing PK parameters measured forPosiphen® and its metabolites in various species.

DETAILED DESCRIPTION OF THE INVENTION

The present invention includes an amount of Compound (1) or a saltthereof that, when administered to a subject, results in a minimum peakplasma circulating level and/or a minimum steady state plasmacirculating level of Compound (1) or a selected metabolite thereof.Dosing of a subject with formulations comprising such amounts of theinvention allow effective levels of the drug and/or any potentiallyactive metabolite to be maintained in circulation in the subject for aspecified period of time, leading to effective inhibition of AChEactivity and/or effective reduction in levels of a neurotoxicaggregating protein in the subject. This circulating level is associatedwith inhibition of synthesis of a neurotoxic aggregating protein, suchas but not limited to APP, Aβ, SNCA, NAC, SOD, HTT, Tau or a prion inthe subject. In one aspect, the compositions and methods of theinvention are thus useful for treating, ameliorating or preventingdementia in the subject. In one embodiment, the dementia is Alzheimer'sdisease, whereby administering the compositions of the inventioninhibits synthesis of APP. In another embodiment the dementia isselected from the group consisting of Parkinson's disease, Huntington'sdisease, Prion's disease, Amyloid Lateral Sclerosis and a Tauopathy.

DEFINITIONS

As used herein, each of the following terms has the meaning associatedwith it in this section.

Unless defined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this invention belongs. Generally,the nomenclature used herein and the laboratory procedures inbiochemistry, analytical chemistry and organic chemistry are thosewell-known and commonly employed in the art. Standard techniques ormodifications thereof are used for chemical syntheses and chemicalanalyses.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e. to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

The term “about” will be understood by persons of ordinary skill in theart and will vary to some extent on the context in which it is used.

As used herein, the terms “Posiphen®” and “Compound (1)” are usedinterchangeably to refer to(3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate or a salt thereof.

As used herein, the terms “N¹-nor-Posiphen” and “Compound (2)” are usedinterchangeably to refer to(3aR)-3a,8-dimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate or a salt thereof.

As used herein, the terms “N⁸-nor-Posiphen” and “Compound (3” are usedinterchangeably to refer to(3aR)-1,3a-dimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate or a salt thereof.

As used herein, the terms “N¹,N⁸-nor-Posiphen” and “Compound (4)” areused interchangeably to refer to(3aR)-3a-methyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate or a salt thereof.

As used herein, the term “APP” refers to amyloid precursor protein.

As used herein, the term “Aβ” refers to amyloid-β peptide.

As used herein, the term “SOD” refers to superoxide dismutase proteinsand members of its family, such as SOD1 and SOD2. The term “SOD”includes any known mutations in superoxide dismutase proteins, such asA4V and G93A mutants in SOD1.

As used herein, the term “SNCA” refers to alpha-synuclein or non-A4component of amyloid precursor.

As use herein, the term “NAC” refers to the alpha-synuclein fragmentknown as the non-Aβ component (NAC) of Alzheimer's disease amyloid.

As used herein, the term “prion” refers to an infectious agent composedof protein in a misfolded form.

As used herein, the term “TSE” refers to any form or variety of atransmissible spongiform encephalopathy.

As used herein, the term “TSE prion” refers to a prion associate with aTSE. Non-limiting examples of TSE prions are scrapie prion;transmissible mink encephalopathy (TME) prion; chronic wasting disease(CWD) prion; bovine spongiform encephalopathy (BSE) prion; felinespongiform encephalopathy (FSE) prion; exotic ungulate encephalopathy(EUE) prion; kuru prion; Creutzfeldt-Jakob disease (CJD) or VariantCreutzfeldt-Jakob disease (vCJD, nvCJD) prion;Gerstmann-Sträussler-Scheinker syndrome (GSS) prion; and fatal familialinsomnia (FFI) prion.

As used herein, the term “HTT” refers to huntingtin or the Huntingtonprotein.

As used herein, the term “Tau” refers to any of the products ofalternative splicing from the gene designated MAPT.

As used herein, the term “neurotoxin aggregating protein” refers to aprotein or family of proteins that has neurotoxic effect uponaccumulating in a tissue of the brain, such as the brain tissue.Non-limiting examples of neurotoxin aggregating proteins are APP, Aβ,SOD, SNCA, NAC, TSE amyloid plaque, HTT, and Tau.

As used herein, the terms “protein”, “peptide” and “polypeptide” areused interchangeably, and refer to a compound comprised of amino acidresidues covalently linked by peptide bonds.

By the term “specifically binds,” as used herein, is meant a molecule,such as an antibody or a small molecule, which recognizes and binds toanother molecule or feature, but does not substantially recognize orbind other molecules or features in a sample.

The phrase “inhibit,” as used herein, means to reduce a molecule, areaction, an interaction, a gene, an mRNA, and/or a protein'sexpression, stability, function or activity by a measurable amount or toprevent entirely. Inhibitors are compounds that, e.g., bind to,partially or totally block stimulation, decrease, prevent, delayactivation, inactivate, desensitize, or down regulate a protein, a gene,and an mRNA stability, expression, function and activity, e.g.,antagonists.

“Effective amount” or “therapeutically effective amount” are usedinterchangeably herein, and refer to an amount of a compound,formulation, material, or composition, as described herein effective toachieve a particular biological result. Such results may include, butare not limited to, the treatment of a disease or condition asdetermined by any means suitable in the art.

As used herein, the term “pharmaceutical composition” refers to amixture of at least one compound of the invention with other chemicalcomponents, such as carriers, stabilizers, diluents, dispersing agents,suspending agents, thickening agents, and/or excipients. Thepharmaceutical composition facilitates administration of the compound toan organism. Multiple techniques of administering a compound exist inthe art including, but not limited to, intravenous, oral, aerosol,parenteral, ophthalmic, pulmonary and topical administration.

“Pharmaceutically acceptable” refers to those properties and/orsubstances that are acceptable to the patient from apharmacological/toxicological point of view and to the manufacturingpharmaceutical chemist from a physical/chemical point of view regardingcomposition, formulation, stability, patient acceptance andbioavailability.

“Pharmaceutically acceptable carrier” refers to a medium that does notinterfere with the effectiveness of the biological activity of theactive ingredient(s) and is not toxic to the host to which it isadministered.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent or encapsulating material,involved in carrying or transporting a compound useful within theinvention within or to the patient such that it may perform its intendedfunction. Typically, such constructs are carried or transported from oneorgan, or portion of the body, to another organ, or portion of the body.Each carrier must be “acceptable” in the sense of being compatible withthe other ingredients of the formulation, including the compound usefulwithin the invention, and not injurious to the patient. Some examples ofmaterials that may serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; surface active agents; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffersolutions; and other non-toxic compatible substances employed inpharmaceutical formulations.

As used herein, “pharmaceutically acceptable carrier” also includes anyand all coatings, antibacterial and antifungal agents, and absorptiondelaying agents, and the like that are compatible with the activity ofthe compound useful within the invention, and are physiologicallyacceptable to the patient. Supplementary active compounds may also beincorporated into the compositions.

As used herein, the “pharmaceutically acceptable carrier” may furtherinclude a pharmaceutically acceptable salt of the compound useful withinthe invention. Other additional ingredients that may be included in thepharmaceutical compositions used in the practice of the invention areknown in the art and described, for example in Remington'sPharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton,Pa.), which is incorporated herein by reference.

As used herein, the term “salt” embraces addition salts of free acids orfree bases that are compounds useful within the invention. Suitable acidaddition salts may be prepared from an inorganic acid or from an organicacid. Examples of inorganic acids include hydrochloric, hydrobromic,hydriodic, nitric, carbonic, sulfuric, phosphoric acids, perchloric andtetrafluoroboronic acids. Appropriate organic acids may be selected fromaliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic,carboxylic and sulfonic classes of organic acids, examples of whichinclude formic, acetic, propionic, succinic, glycolic, gluconic, lactic,malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic,aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic,phenylacetic, mandelic, embonic (pamoic), methanesulfonic,ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic,2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic,galactaric and galacturonic acid. Suitable base addition salts ofcompounds useful within the invention include, for example, metallicsalts including alkali metal, alkaline earth metal and transition metalsalts such as, for example, lithium, calcium, magnesium, potassium,sodium and zinc salts. Acceptable base addition salts also includeorganic salts made from basic amines such as, for example,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methyl-glutamine) and procaine. All ofthese salts may be prepared by conventional means from the correspondingfree base compound by reacting, for example, the appropriate acid orbase with the corresponding free base.

An “individual”, “patient” or “subject”, as that term is used herein,includes a member of any animal species including, but are not limitedto, birds, humans and other primates, and other mammals includingcommercially relevant mammals such as cattle, pigs, horses, sheep, cats,and dogs. Preferably, the subject is a human.

The term “treat” or “treating”, as used herein, means reducing thefrequency with which symptoms are experienced by a subject oradministering an agent or compound to reduce the frequency and/orseverity with which symptoms are experienced. As used herein,“alleviate” is used interchangeably with the term “treat.” Treating adisease, disorder or condition may or may not include completeeradication or elimination of the symptom. The term “therapeutic” asused herein means a treatment and/or prophylaxis

“Instructional material,” as that term is used herein, includes apublication, a recording, a diagram, or any other medium of expressionthat can be used to communicate the usefulness of the composition and/orcompound of the invention in a kit. The instructional material of thekit may for example, be affixed to a container that contains thecompound and/or composition of the invention or be shipped together witha container which contains the compound and/or composition.Alternatively, the instructional material may be shipped separately fromthe container with the intention that the recipient uses theinstructional material and the compound cooperatively. Delivery of theinstructional material may be, for example, by physical delivery of thepublication or other medium of expression communicating the usefulnessof the kit, or may alternatively be achieved by electronic transmission,for example by means of a computer, such as by electronic mail, ordownload from a website.

Throughout this disclosure, various aspects of the invention can bepresented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible sub-ranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. Thisapplies regardless of the breadth of the range.

Compounds and Compositions of the Invention

In one aspect, the compound useful within the methods of the inventionis Compound (1):

or a salt thereof.

Compound (1) or any other compound useful within the methods of theinvention may be synthesized using techniques well-known in the art oforganic synthesis or obtained from commercial sources. In a non-limitingexample, Compound (1) may be synthesized according to the disclosure inU.S. Pat. No. 6,495,700, incorporated herein in its entirety.

The invention includes an amount of Compound (1) or a salt thereof,wherein administering the amount to a subject results in a peak plasmacirculating level of Compound (1) ranging from about 10 ng/ml to about160 ng/mL in the subject.

In one embodiment, the peak plasma circulating level ranges from about80 ng/mL to about 160 ng/ml in the subject. In another embodiment, thepeak plasma circulating level is reached within about 6 hours after theadministering. In yet another embodiment, the peak plasma circulatinglevel is reached within about 3 hours after the administering. In yetanother embodiment, the plasma circulating level of Compound (1) isequal to or greater than about 20 ng/mL for at least 12 hours after theadministering. In yet another embodiment, the plasma circulating levelof Compound (1) is equal to or greater than about 20 ng/mL for at least9 hours after the administering. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (2)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (3)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (4)ranging from about 1% to about 9% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a steady state plasma concentration of Compound(1) of at least about 100 ng/ml in the subject. In yet anotherembodiment, the administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in the subject.In yet another embodiment, the administering results in a steady stateplasma concentration of Compound (3) of at least about 10 ng/ml in thesubject. In yet another embodiment, the administering results in asteady state plasma concentration of Compound (4) of at least about 3ng/ml in the subject. In yet another embodiment, the administeringresults in a brain level of Compound (1) that ranges from about 4 toabout 10 times the plasma level of Compound (1) in the subject. In yetanother embodiment, the brain level of Compound (2) ranges from about15% to about 150% of the brain level of Compound (1) in the subject. Inyet another embodiment, the brain level of Compound (3) ranges fromabout 15% to about 150% of the brain level of Compound (1) in thesubject. In yet another embodiment, the brain level of Compound (4) islower than the brain level of Compound (2) or Compound (3) in thesubject. In yet another embodiment, the subject is a human. In yetanother embodiment, the peak plasma circulating level of Compound (1) islower than the peak toxic level of Compound (1) in the plasma of thesubject. In a non-limiting embodiment, the toxic level of Compound (1)in the plasma is greater than 80 ng/mL.

Salts of the Compounds of the Invention

The compounds described herein may form salts with acids or bases, andsuch salts are included in the present invention. In one embodiment, thesalts are pharmaceutically acceptable salts. The term “salts” embracesaddition salts of free acids or free bases that are compounds of theinvention. The term “pharmaceutically acceptable salt” refers to saltsthat possess toxicity profiles within a range that affords utility inpharmaceutical applications. Pharmaceutically unacceptable salts maynonetheless possess properties such as high crystallinity, which haveutility in the practice of the present invention, such as for exampleutility in process of synthesis, purification or formulation ofcompounds of the invention.

Suitable pharmaceutically acceptable acid addition salts may be preparedfrom an inorganic acid or from an organic acid. Examples of inorganicacids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic,sulfuric, and phosphoric acids. Appropriate organic acids may beselected from aliphatic, cycloaliphatic, aromatic, araliphatic,heterocyclic, carboxylic and sulfonic classes of organic acids, examplesof which include formic, acetic, propionic, succinic, glycolic,gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic,fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic,4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic,sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric,salicylic, galactaric and galacturonic acid.

Examples of pharmaceutically unacceptable acid addition salts include,for example, perchlorates and tetrafluoroborates.

Suitable pharmaceutically acceptable base addition salts of compounds ofthe invention include, for example, metallic salts including alkalimetal, alkaline earth metal and transition metal salts such as, forexample, calcium, magnesium, potassium, sodium and zinc salts.Pharmaceutically acceptable base addition salts also include organicsalts made from basic amines such as, for example,N,N′-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methylglucamine) and procaine. Examples ofpharmaceutically unacceptable base addition salts include lithium saltsand cyanate salts. All of these salts may be prepared from thecorresponding compound by reacting, for example, the appropriate acid orbase

Methods of the Invention

The invention includes a method of inhibiting production of a neurotoxicaggregating protein, such as APP, Aβ, SNCA, NAC, SOD, Tau, TSE prion andHTT in a subject. The method comprises administering to the subject apharmaceutical composition comprising a pharmaceutically acceptablecarrier and an amount of Compound (1) or a salt thereof, whereinadministering the composition to the subject results in a peak plasmacirculating level of Compound (1) ranging from about 10 ng/ml to about160 ng/ml in the subject, whereby the production of the neurotoxicaggregating protein in the subject is inhibited.

The invention includes a method of treating dementia in a subject. Themethod comprising administering to the subject a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier and anamount of Compound (1) or a salt thereof, wherein administering thecomposition to the subject results in a peak plasma circulating level ofCompound (1) ranging from about 10 ng/ml to about 160 ng/ml in thesubject, whereby the dementia in the subject is treated.

In one embodiment, the peak plasma circulating level ranges from about80 ng/mL to about 160 ng/ml in the subject. In another embodiment, thepeak plasma circulating level is reached within about 6 hours after theadministering. In yet another embodiment, the peak plasma circulatinglevel is reached within about 3 hours after the administering. In yetanother embodiment, the plasma circulating level of Compound (1) isequal to or greater than about 20 ng/mL for at least 12 hours after theadministering. In yet another embodiment, the plasma circulating levelof Compound (1) is equal to or greater than about 20 ng/mL for at least9 hours after the administering. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (2)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (3)ranging from about 15% to about 30% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a peak plasma concentration of Compound (4)ranging from about 1% to about 9% of the peak plasma concentration ofCompound (1) in the subject. In yet another embodiment, theadministering results in a steady state plasma concentration of Compound(1) of at least about 100 ng/ml in the subject. In yet anotherembodiment, the administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in the subject.In yet another embodiment, the administering results in a steady stateplasma concentration of Compound (3) of at least about 10 ng/ml in thesubject. In yet another embodiment, the administering results in asteady state plasma concentration of Compound (4) of at least about 3ng/ml in the subject. In yet another embodiment, the administeringresults in a brain level of Compound (1) that ranges from about 4 toabout 10 times the plasma level of Compound (1) in the subject. In yetanother embodiment, the brain level of Compound (2) ranges from about15% to about 150% of the brain level of Compound (1) in the subject. Inyet another embodiment, the brain level of Compound (3) ranges fromabout 15% to about 150% of the brain level of Compound (1) in thesubject. In yet another embodiment, the brain level of Compound (4) islower than the brain level of Compound (2) or Compound (3) in thesubject. In yet another embodiment, the subject is a human. In yetanother embodiment, the peak plasma circulating level of Compound (1) islower than the peak toxic level of Compound (1) in the plasma of thesubject. In a non-limiting embodiment, the toxic level of Compound (1)in the plasma is greater than 80 ng/mL.

In one embodiment, the administering results in a reduction of orgreater than about 25% in a cerebrospinal fluid marker selected from thegroup consisting of sAPPα, sAPPβ, Tau and pTau in the subject. Inanother embodiment, the administering results in a reduction of orgreater than about 30% in a cerebrospinal fluid marker selected from thegroup consisting of sAPPα, sAPPβ, Tau and pTau in the subject. In yetanother embodiment, the subject is a human. In yet another embodiment,the dementia is Alzheimer's disease. In yet another embodiment, thedementia is selected from the group consisting of Parkinson's disease,Huntington's disease, Prion's disease, Amyloid Lateral Sclerosis and atauopathy.

Pharmaceutical Compositions and Therapies Administration of a compounduseful within the invention may be achieved in a number of differentways, using methods known in the art. The therapeutic and prophylacticmethods of the invention thus encompass the use of pharmaceuticalcompositions comprising the compounds useful within the invention topractice the methods of the invention. The pharmaceutical compositionsuseful for practicing the invention may be administered to deliver adose of 1 ng/kg/day to 100 mg/kg/day.

The relative amounts of the active ingredient, the pharmaceuticallyacceptable carrier, and any additional ingredients in a pharmaceuticalcomposition of the invention will vary, depending upon the identity,size, and condition of the subject treated and further depending uponthe route by which the composition is to be administered. By way ofexample, the composition may comprise between 0.1% and 100% (w/w) activeingredient.

Although the description of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions that aresuitable for ethical administration to humans, it will be understood bythe skilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and perform such modification with merely ordinary, if any,experimentation. Subjects to which administration of the pharmaceuticalcompositions of the invention is contemplated include, but are notlimited to, humans and other primates, mammals including commerciallyrelevant mammals such as non-human primates, cattle, pigs, horses,sheep, cats, and dogs.

Typically, dosages which may be administered in a method of theinvention to an animal, preferably a human, range in amount from 0.5 μgto about 50 mg per kilogram of body weight of the animal. While theprecise dosage administered will vary depending upon any number offactors, including but not limited to, the type of animal and type ofdisease state being treated, the age of the animal and the route ofadministration, the dosage of the compound will preferably vary fromabout 1 μg to about 10 mg per kilogram of body weight of the animal.More preferably, the dosage will vary from about 3 μg to about 1 mg perkilogram of body weight of the animal.

Pharmaceutical compositions that are useful in the methods of theinvention may be prepared, packaged, or sold in formulations suitablefor oral, parenteral, topical, buccal, or another route ofadministration. Other contemplated formulations include projectednanoparticles, liposomal preparations, resealed erythrocytes containingthe active ingredient, and immunologically-based formulations.

The formulations of the pharmaceutical compositions described herein maybe prepared by any method known or hereafter developed in the art ofpharmacology. In general, such preparatory methods include the step ofbringing the active ingredient into association with a carrier or one ormore other accessory ingredients, and then, if necessary or desirable,shaping or packaging the product into a desired single- or multi-doseunit.

A pharmaceutical composition of the invention may be prepared, packaged,or sold in bulk, as a single unit dose, or as a plurality of single unitdoses. As used herein, a “unit dose” is discrete amount of thepharmaceutical composition comprising a predetermined amount of theactive ingredient. The amount of the active ingredient is generallyequal to the dosage of the active ingredient which would be administeredto a subject or a convenient fraction of such a dosage such as, forexample, one-half or one-third of such a dosage.

In one embodiment, the compositions of the invention are formulatedusing one or more pharmaceutically acceptable excipients or carriers. Inone embodiment, the pharmaceutical compositions of the inventioncomprise a therapeutically effective amount of a compound or conjugateof the invention and a pharmaceutically acceptable carrier.Pharmaceutically acceptable carriers that are useful, include, but arenot limited to, glycerol, water, saline, ethanol and otherpharmaceutically acceptable salt solutions such as phosphates and saltsof organic acids. Examples of these and other pharmaceuticallyacceptable carriers are described in Remington's Pharmaceutical Sciences(1991, Mack Publication Co., New Jersey).

The carrier may be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetable oils. The proper fluidity may be maintained, forexample, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms may be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol,in the composition. Prolonged absorption of the injectable compositionsmay be brought about by including in the composition an agent thatdelays absorption, for example, aluminum monostearate or gelatin. In oneembodiment, the pharmaceutically acceptable carrier is not DMSO alone.

Formulations may be employed in admixtures with conventional excipients,i.e., pharmaceutically acceptable organic or inorganic carriersubstances suitable for oral, vaginal, parenteral, nasal, intravenous,subcutaneous, enteral, or any other suitable mode of administration,known to the art. The pharmaceutical preparations may be sterilized andif desired mixed with auxiliary agents, e.g., lubricants, preservatives,stabilizers, wetting agents, emulsifiers, salts for influencing osmoticpressure buffers, coloring, flavoring and/or aromatic substances and thelike. They may also be combined where desired with other active agents,e.g., other analgesic agents.

As used herein, “additional ingredients” include, but are not limitedto, one or more of the following: excipients; surface active agents;dispersing agents; inert diluents; granulating and disintegratingagents; binding agents; lubricating agents; sweetening agents; flavoringagents; coloring agents; preservatives; physiologically degradablecompositions such as gelatin; aqueous vehicles and solvents; oilyvehicles and solvents; suspending agents; dispersing or wetting agents;emulsifying agents, demulcents; buffers; salts; thickening agents;fillers; emulsifying agents; antioxidants; antibiotics; antifungalagents; stabilizing agents; and pharmaceutically acceptable polymeric orhydrophobic materials. Other “additional ingredients” that may beincluded in the pharmaceutical compositions of the invention are knownin the art and described, for example in Genaro, ed. (1985, Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa.), which isincorporated herein by reference.

The composition of the invention may comprise a preservative from about0.005% to 2.0% by total weight of the composition. The preservative isused to prevent spoilage in the case of exposure to contaminants in theenvironment. Examples of preservatives useful in accordance with theinvention included but are not limited to those selected from the groupconsisting of benzyl alcohol, sorbic acid, parabens, imidurea andcombinations thereof. A particularly preferred preservative is acombination of about 0.5% to 2.0% benzyl alcohol and 0.05% to 0.5%sorbic acid.

The composition preferably includes an anti-oxidant and a chelatingagent that inhibits the degradation of the compound. Preferredantioxidants for some compounds are BHT, BHA, alpha-tocopherol andascorbic acid in the preferred range of about 0.01% to 0.3% and morepreferably BHT in the range of 0.03% to 0.1% by weight by total weightof the composition. Preferably, the chelating agent is present in anamount of from 0.01% to 0.5% by weight by total weight of thecomposition. Particularly preferred chelating agents include edetatesalts (e.g. disodium edetate) and citric acid in the weight range ofabout 0.01% to 0.20% and more preferably in the range of 0.02% to 0.10%by weight by total weight of the composition. The chelating agent isuseful for chelating metal ions in the composition that may bedetrimental to the shelf life of the formulation. While BHT and disodiumedetate are the particularly preferred antioxidant and chelating agentrespectively for some compounds, other suitable and equivalentantioxidants and chelating agents may be substituted therefore as wouldbe known to those skilled in the art.

Liquid suspensions may be prepared using conventional methods to achievesuspension of the active ingredient in an aqueous or oily vehicle.Aqueous vehicles include, for example, water, and isotonic saline. Oilyvehicles include, for example, almond oil, oily esters, ethyl alcohol,vegetable oils such as arachis, olive, sesame, or coconut oil,fractionated vegetable oils, and mineral oils such as liquid paraffin.Liquid suspensions may further comprise one or more additionalingredients including, but not limited to, suspending agents, dispersingor wetting agents, emulsifying agents, demulcents, preservatives,buffers, salts, flavorings, coloring agents, and sweetening agents. Oilysuspensions may further comprise a thickening agent. Known suspendingagents include, but are not limited to, sorbitol syrup, hydrogenatededible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gumacacia, and cellulose derivatives such as sodium carboxymethylcellulose,methylcellulose, hydroxypropylmethylcellulose. Known dispersing orwetting agents include, but are not limited to, naturally-occurringphosphatides such as lecithin, condensation products of an alkyleneoxide with a fatty acid, with a long chain aliphatic alcohol, with apartial ester derived from a fatty acid and a hexitol, or with a partialester derived from a fatty acid and a hexitol anhydride (e.g.,polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylenesorbitol monooleate, and polyoxyethylene sorbitan monooleate,respectively). Known emulsifying agents include, but are not limited to,lecithin, and acacia. Known preservatives include, but are not limitedto, methyl, ethyl, or n-propyl-para-hydroxybenzoates, ascorbic acid, andsorbic acid. Known sweetening agents include, for example, glycerol,propylene glycol, sorbitol, sucrose, and saccharin. Known thickeningagents for oily suspensions include, for example, beeswax, hardparaffin, and cetyl alcohol.

Liquid solutions of the active ingredient in aqueous or oily solventsmay be prepared in substantially the same manner as liquid suspensions,the primary difference being that the active ingredient is dissolved,rather than suspended in the solvent. As used herein, an “oily” liquidis one which comprises a carbon-containing liquid molecule and whichexhibits a less polar character than water. Liquid solutions of thepharmaceutical composition of the invention may comprise each of thecomponents described with regard to liquid suspensions, it beingunderstood that suspending agents will not necessarily aid dissolutionof the active ingredient in the solvent. Aqueous solvents include, forexample, water, and isotonic saline. Oily solvents include, for example,almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis,olive, sesame, or coconut oil, fractionated vegetable oils, and mineraloils such as liquid paraffin.

Powdered and granular formulations of a pharmaceutical preparation ofthe invention may be prepared using known methods. Such formulations maybe administered directly to a subject, used, for example, to formtablets, to fill capsules, or to prepare an aqueous or oily suspensionor solution by addition of an aqueous or oily vehicle thereto. Each ofthese formulations may further comprise one or more of dispersing orwetting agent, a suspending agent, and a preservative. Additionalexcipients, such as fillers and sweetening, flavoring, or coloringagents, may also be included in these formulations.

A pharmaceutical composition of the invention may also be prepared,packaged, or sold in the form of oil-in-water emulsion or a water-in-oilemulsion. The oily phase may be a vegetable oil such as olive or arachisoil, a mineral oil such as liquid paraffin, or a combination of these.Such compositions may further comprise one or more emulsifying agentssuch as naturally occurring gums such as gum acacia or gum tragacanth,naturally-occurring phosphatides such as soybean or lecithinphosphatide, esters or partial esters derived from combinations of fattyacids and hexitol anhydrides such as sorbitan monooleate, andcondensation products of such partial esters with ethylene oxide such aspolyoxyethylene sorbitan monooleate. These emulsions may also containadditional ingredients including, for example, sweetening or flavoringagents.

Methods for impregnating or coating a material with a chemicalcomposition are known in the art, and include, but are not limited tomethods of depositing or binding a chemical composition onto a surface,methods of incorporating a chemical composition into the structure of amaterial during the synthesis of the material (i.e., such as with aphysiologically degradable material), and methods of absorbing anaqueous or oily solution or suspension into an absorbent material, withor without subsequent drying.

Controlled- or sustained-release formulations of a composition of theinvention may be made using conventional technology, in addition to thedisclosure set forth elsewhere herein. In some cases, the dosage formsto be used can be provided as slow or controlled-release of one or moreactive ingredients therein using, for example, hydropropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmoticsystems, multilayer coatings, microparticles, liposomes, or microspheresor a combination thereof to provide the desired release profile invarying proportions. Suitable controlled-release formulations known tothose of ordinary skill in the art, including those described herein,can be readily selected for use with the compositions of the invention.

Controlled-release of an active ingredient can be stimulated by variousinducers, for example pH, temperature, enzymes, water, or otherphysiological conditions or compounds. The term “controlled-releasecomponent” in the context of the present invention is defined herein asa compound or compounds, including, but not limited to, polymers,polymer matrices, gels, permeable membranes, liposomes, nanoparticles,or microspheres or a combination thereof that facilitates thecontrolled-release of the active ingredient.

Administration/Dosing

The regimen of administration may affect what constitutes an effectiveamount. The therapeutic formulations may be administered to the subjecteither prior to or after a diagnosis of disease. Further, severaldivided dosages, as well as staggered dosages may be administered dailyor sequentially, or the dose may be continuously infused, or may be abolus injection. Further, the dosages of the therapeutic formulationsmay be proportionally increased or decreased as indicated by theexigencies of the therapeutic or prophylactic situation.

Administration of the compositions of the present invention to asubject, preferably a mammal, more preferably a human, may be carriedout using known procedures, at dosages and for periods of time effectiveto prevent or treat disease. An effective amount of the therapeuticcompound necessary to achieve a therapeutic effect may vary according tofactors such as the activity of the particular compound employed; thetime of administration; the rate of excretion of the compound; theduration of the treatment; other drugs, compounds or materials used incombination with the compound; the state of the disease or disorder,age, sex, weight, condition, general health and prior medical history ofthe subject being treated, and like factors well-known in the medicalarts. Dosage regimens may be adjusted to provide the optimum therapeuticresponse. For example, several divided doses may be administered dailyor the dose may be proportionally reduced as indicated by the exigenciesof the therapeutic situation. A non-limiting example of an effectivedose range for a therapeutic compound of the invention is from about 1and 5,000 mg/kg of body weight/per day. One of ordinary skill in the artwould be able to study the relevant factors and make the determinationregarding the effective amount of the therapeutic compound without undueexperimentation.

The compound may be administered to an animal as frequently as severaltimes daily, or it may be administered less frequently, such as once aday, once a week, once every two weeks, once a month, or even lessfrequently, such as once every several months or even once a year orless. The frequency of the dose will be readily apparent to the skilledartisan and will depend upon any number of factors, such as, but notlimited to, the type and severity of the disease being treated, the typeand age of the animal, etc. The formulations of the pharmaceuticalcompositions described herein may be prepared by any method known orhereafter developed in the art of pharmacology. In general, suchpreparatory methods include the step of bringing the active ingredientinto association with a carrier or one or more other accessoryingredients, and then, if necessary or desirable, shaping or packagingthe product into a desired single- or multi-dose unit.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular subject, composition, and mode ofadministration, without being toxic to the subject.

A medical doctor, e.g., physician or veterinarian, having ordinary skillin the art may readily determine and prescribe the effective amount ofthe pharmaceutical composition required. For example, the physician orveterinarian could start doses of the compounds of the inventionemployed in the pharmaceutical composition at levels lower than thatrequired in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulatethe compound in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subjects tobe treated; each unit containing a predetermined quantity of therapeuticcompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical vehicle. The dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding/formulating such a therapeutic compound for thetreatment of a disease in a subject.

In one embodiment, the compositions of the invention are administered tothe subject in dosages that range from one to five times per day ormore. In another embodiment, the compositions of the invention areadministered to the subject in range of dosages that include, but arenot limited to, once every day, every two, days, every three days toonce a week, and once every two weeks. It will be readily apparent toone skilled in the art that the frequency of administration of thevarious combination compositions of the invention will vary from subjectto subject depending on many factors including, but not limited to, age,disease or disorder to be treated, gender, overall health, and otherfactors. Thus, the invention should not be construed to be limited toany particular dosage regime and the precise dosage and composition tobe administered to any subject will be determined by the attendingphysical taking all other factors about the subject into account.

Compounds of the invention for administration may be in the range offrom about 1 mg to about 1,000 mg, about 2 mg to about 950 mg, about 4mg to about 900 mg, about 7.5 mg to about 850 mg, about 15 mg to about750 mg, about 20 mg to about 700 mg, about 30 mg to about 600 mg, about50 mg to about 500 mg, about 75 mg to about 400 mg, about 100 mg toabout 300 mg, about 120 mg to about 250 mg, and any and all whole orpartial increments therebetween.

In some embodiments, the dose of a compound of the invention is fromabout 1 mg and about 2,500 mg. In some embodiments, a dose of a compoundof the invention used in compositions described herein is less thanabout 10,000 mg, or less than about 8,000 mg, or less than about 6,000mg, or less than about 5,000 mg, or less than about 3,000 mg, or lessthan about 2,000 mg, or less than about 1,000 mg, or less than about 500mg, or less than about 200 mg, or less than about 50 mg. Similarly, insome embodiments, a dose of a second compound (i.e., a drug used fortreating the same or another disease as that treated by the compositionsof the invention) as described herein is less than about 1,000 mg, orless than about 800 mg, or less than about 600 mg, or less than about500 mg, or less than about 400 mg, or less than about 300 mg, or lessthan about 200 mg, or less than about 100 mg, or less than about 50 mg,or less than about 40 mg, or less than about 30 mg, or less than about25 mg, or less than about 20 mg, or less than about 15 mg, or less thanabout 10 mg, or less than about 5 mg, or less than about 2 mg, or lessthan about 1 mg, or less than about 0.5 mg, and any and all whole orpartial increments thereof.

In one embodiment, the present invention is directed to a packagedpharmaceutical composition comprising a container holding atherapeutically effective amount of a composition of the invention,alone or in combination with a second pharmaceutical agent; andinstructions for using the composition to treat, prevent, or reduce oneor more symptoms of a disease in a subject.

Routes of Administration

Routes of administration of any of the compositions of the inventioninclude oral, nasal, rectal, parenteral, sublingual, transdermal,transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral,vaginal (e.g., trans- and perivaginally), (intra)nasal, and(trans)rectal), intravesical, intrapulmonary, intraduodenal,intragastrical, intrathecal, subcutaneous, intramuscular, intradermal,intra-arterial, intravenous, intrabronchial, inhalation, and topicaladministration.

Suitable compositions and dosage forms include, for example, tablets,capsules, caplets, pills, gel caps, troches, dispersions, suspensions,solutions, syrups, granules, beads, transdermal patches, gels, powders,pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs,suppositories, liquid sprays for nasal or oral administration, drypowder or aerosolized formulations for inhalation, compositions andformulations for intravesical administration and the like. It should beunderstood that the formulations and compositions that would be usefulin the present invention are not limited to the particular formulationsand compositions that are described herein.

Oral Administration

For oral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules, caplets and gelcaps. Otherformulations suitable for oral administration include, but are notlimited to, a powdered or granular formulation, an aqueous or oilysuspension, an aqueous or oily solution, a paste, a gel, toothpaste, amouthwash, a coating, an oral rinse, or an emulsion. The compositionsintended for oral use may be prepared according to any method known inthe art and such compositions may contain one or more agents selectedfrom the group consisting of inert, non-toxic pharmaceuticallyexcipients that are suitable for the manufacture of tablets. Suchexcipients include, for example an inert diluent such as lactose;granulating and disintegrating agents such as cornstarch; binding agentssuch as starch; and lubricating agents such as magnesium stearate.

Tablets may be non-coated or they may be coated using known methods toachieve delayed disintegration in the gastrointestinal tract of asubject, thereby providing sustained release and absorption of theactive ingredient. By way of example, a material such as glycerylmonostearate or glyceryl distearate may be used to coat tablets. Furtherby way of example, tablets may be coated using methods described in U.S.Pat. Nos. 4,256,108; 4,160,452; and 4,265,874 to form osmoticallycontrolled release tablets. Tablets may further comprise a sweeteningagent, a flavoring agent, a coloring agent, a preservative, or somecombination of these in order to provide for pharmaceutically elegantand palatable preparation.

Hard capsules comprising the active ingredient may be made using aphysiologically degradable composition, such as gelatin. Such hardcapsules comprise the active ingredient, and may further compriseadditional ingredients including, for example, an inert solid diluentsuch as calcium carbonate, calcium phosphate, or kaolin.

Soft gelatin capsules comprising the active ingredient may be made usinga physiologically degradable composition, such as gelatin. Such softcapsules comprise the active ingredient, which may be mixed with wateror an oil medium such as peanut oil, liquid paraffin, or olive oil.

For oral administration, the compositions of the invention may be in theform of tablets or capsules prepared by conventional means withpharmaceutically acceptable excipients such as binding agents; fillers;lubricants; disintegrates; or wetting agents. If desired, the tabletsmay be coated using suitable methods and coating materials such asOPADRY® film coating systems available from Colorcon, West Point, Pa.(e.g., OPADRY® OY Type, OYC Type, Organic Enteric OY-P Type, AqueousEnteric OY-A Type, OY-PM Type and OPADRY® White, 32K18400).

Liquid preparation for oral administration may be in the form ofsolutions, syrups or suspensions. The liquid preparations may beprepared by conventional means with pharmaceutically acceptableadditives such as suspending agents (e.g., sorbitol syrup, methylcellulose or hydrogenated edible fats); emulsifying agent (e.g.,lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily estersor ethyl alcohol); and preservatives (e.g., methyl or propylpara-hydroxy benzoates or sorbic acid). Liquid formulations of apharmaceutical composition of the invention which are suitable for oraladministration may be prepared, packaged, and sold either in liquid formor in the form of a dry product intended for reconstitution with wateror another suitable vehicle prior to use.

A tablet comprising the active ingredient may, for example, be made bycompressing or molding the active ingredient, optionally with one ormore additional ingredients. Compressed tablets may be prepared bycompressing, in a suitable device, the active ingredient in afree-flowing form such as a powder or granular preparation, optionallymixed with one or more of a binder, a lubricant, an excipient, a surfaceactive agent, and a dispersing agent. Molded tablets may be made bymolding, in a suitable device, a mixture of the active ingredient, apharmaceutically acceptable carrier, and at least sufficient liquid tomoisten the mixture. Pharmaceutically acceptable excipients used in themanufacture of tablets include, but are not limited to, inert diluents,granulating and disintegrating agents, binding agents, and lubricatingagents. Known dispersing agents include, but are not limited to, potatostarch and sodium starch glycollate. Known surface-active agentsinclude, but are not limited to, sodium lauryl sulphate. Known diluentsinclude, but are not limited to, calcium carbonate, sodium carbonate,lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogenphosphate, and sodium phosphate. Known granulating and disintegratingagents include, but are not limited to, corn starch and alginic acid.Known binding agents include, but are not limited to, gelatin, acacia,pre-gelatinized maize starch, polyvinylpyrrolidone, and hydroxypropylmethylcellulose. Known lubricating agents include, but are not limitedto, magnesium stearate, stearic acid, silica, and talc.

Granulating techniques are well known in the pharmaceutical art formodifying starting powders or other particulate materials of an activeingredient. The powders are typically mixed with a binder material intolarger permanent free-flowing agglomerates or granules referred to as a“granulation.” For example, solvent-using “wet” granulation processesare generally characterized in that the powders are combined with abinder material and moistened with water or an organic solvent underconditions resulting in the formation of a wet granulated mass fromwhich the solvent must then be evaporated.

Melt granulation generally consists in the use of materials that aresolid or semi-solid at room temperature (i.e. having a relatively lowsoftening or melting point range) to promote granulation of powdered orother materials, essentially in the absence of added water or otherliquid solvents. The low melting solids, when heated to a temperature inthe melting point range, liquefy to act as a binder or granulatingmedium. The liquefied solid spreads itself over the surface of powderedmaterials with which it is contacted, and on cooling, forms a solidgranulated mass in which the initial materials are bound together. Theresulting melt granulation may then be provided to a tablet press or beencapsulated for preparing the oral dosage form. Melt granulationimproves the dissolution rate and bioavailability of an active (i.e.drug) by forming a solid dispersion or solid solution.

U.S. Pat. No. 5,169,645 discloses directly compressible wax-containinggranules having improved flow properties. The granules are obtained whenwaxes are admixed in the melt with certain flow improving additives,followed by cooling and granulation of the admixture. In certainembodiments, only the wax itself melts in the melt combination of thewax(es) and additives(s), and in other cases both the wax(es) and theadditives(s) will melt.

The present invention also includes a multi-layer tablet comprising alayer providing for the delayed release of one or more compounds of theinvention, and a further layer providing for the immediate release of amedication for treatment of a disease. Using a wax/pH-sensitive polymermix, a gastric insoluble composition may be obtained in which the activeingredient is entrapped, ensuring its delayed release.

Parenteral Administration

As used herein, “parenteral administration” of a pharmaceuticalcomposition includes any route of administration characterized byphysical breaching of a tissue of a subject and administration of thepharmaceutical composition through the breach in the tissue. Parenteraladministration thus includes, but is not limited to, administration of apharmaceutical composition by injection of the composition, byapplication of the composition through a surgical incision, byapplication of the composition through a tissue-penetrating non-surgicalwound, and the like. In particular, parenteral administration iscontemplated to include, but is not limited to, intraocular,intravitreal, subcutaneous, intraperitoneal, intramuscular, intrasternalinjection, intratumoral, and kidney dialytic infusion techniques.

Formulations of a pharmaceutical composition suitable for parenteraladministration comprise the active ingredient combined with apharmaceutically acceptable carrier, such as sterile water or sterileisotonic saline. Such formulations may be prepared, packaged, or sold ina form suitable for bolus administration or for continuousadministration. Injectable formulations may be prepared, packaged, orsold in unit dosage form, such as in ampules or in multi-dose containerscontaining a preservative. Formulations for parenteral administrationinclude, but are not limited to, suspensions, solutions, emulsions inoily or aqueous vehicles, pastes, and implantable sustained-release orbiodegradable formulations. Such formulations may further comprise oneor more additional ingredients including, but not limited to,suspending, stabilizing, or dispersing agents. In one embodiment of aformulation for parenteral administration, the active ingredient isprovided in dry (i.e. powder or granular) form for reconstitution with asuitable vehicle (e.g. sterile pyrogen-free water) prior to parenteraladministration of the reconstituted composition.

The pharmaceutical compositions may be prepared, packaged, or sold inthe form of a sterile injectable aqueous or oily suspension or solution.This suspension or solution may be formulated according to the knownart, and may comprise, in addition to the active ingredient, additionalingredients such as the dispersing agents, wetting agents, or suspendingagents described herein. Such sterile injectable formulations may beprepared using a non-toxic parenterally-acceptable diluent or solvent,such as water or 1,3-butanediol, for example. Other acceptable diluentsand solvents include, but are not limited to, Ringer's solution,isotonic sodium chloride solution, and fixed oils such as syntheticmono- or di-glycerides. Other parentally-administrable formulations thatare useful include those which comprise the active ingredient inmicrocrystalline form, in a liposomal preparation, or as a component ofa biodegradable polymer systems. Compositions for sustained release orimplantation may comprise pharmaceutically acceptable polymeric orhydrophobic materials such as an emulsion, an ion exchange resin, asparingly soluble polymer, or a sparingly soluble salt.

Topical Administration

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for topical administration. There areseveral advantages to delivering compounds, including drugs or othertherapeutic agents, into the skin (dermal drug delivery) or into thebody through the skin (transdermal drug delivery). Transdermal compounddelivery offers an attractive alternative to injections and oralmedications. Dermal compound delivery offers an efficient way to delivera compound to the skin of a mammal, and preferably a human, and providesa method of treatment of the skin, or otherwise provides a method ofaffecting the skin, without the need to break or damage the outer layerof the skin. In the present invention, dermal delivery, by way of adermally-acting compound of the invention, provides these advantages fortreatment of a skin-related condition, disorder or disease.

A number of compounds, including some drugs, will penetrate the skineffectively simply because the molecules are relatively small and potentat small doses of 0.1 mg to 15 mg/day (Kanikkannan et al., 2000, Curr.Med. Chem. 7:593-608). Many other compounds and drugs can be deliveredonly when an additional enhancement system is provided to “force” themto pass through the skin. Among several methods of transdermal drugdelivery are electroporation, sonophoresis, iontophoresis, permeationenhancers (cyclodextrins), and liposomes. While the aforementionedmethods are also included in the present invention for dermal deliveryof the compounds of the invention, liposomes represent a preferreddermal delivery method.

The composition of the invention may consist of the active ingredientalone, in a form suitable for administration to a subject, or thecomposition may comprise at least one active ingredient and one or morepharmaceutically acceptable carriers, one or more additionalingredients, or some combination of these. The active ingredient may bepresent in the composition in the form of a physiologically acceptableester or salt, such as in combination with a physiologically acceptablecation or anion, as is well known in the art. Compositions of theinvention will also be understood to encompass pharmaceuticalcompositions useful for treatment of other conditions, disorders anddiseases associated with the skin.

In one aspect, a dermal delivery vehicle of the invention is acomposition comprising at least one first compound that can facilitatedermal delivery of at least one second compound associated with, or inclose physical proximity to, the composition comprising the firstcompound. As will be understood by the skilled artisan, when armed withthe disclosure set forth herein, such delivery vehicles include, butshould not be limited to, liposomes, nanosomes, phospholipid-basednon-liposome compositions (eg., selected cochleates), among others.

Formulations suitable for topical administration include, but are notlimited to, liquid or semi-liquid preparations such as liniments,lotions, oil-in-water or water-in-oil emulsions such as creams,ointments or pastes, and solutions or suspensions.Topically-administrable formulations may, for example, comprise fromabout 0.001% to about 90% (w/w) active ingredient, although theconcentration of the active ingredient may be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

In one aspect of the invention, a dermal delivery system includes aliposome delivery system, and that the present invention should not beconstrued to be limited to any particular liposome delivery system.Based on the disclosure set forth herein, the skilled artisan willunderstand how to identify a liposome delivery system as being useful inthe present invention.

The present invention also encompasses the improvement of dermal andtransdermal drug delivery through the use of penetration enhancers (alsocalled sorption promoters or accelerants), which penetrate into skin toreversibly decrease the barrier resistance. Many compounds are known inthe art for penetration enhancing activity, including sulphoxides (suchas dimethylsulphoxide, DMSO), azones (e.g. laurocapram), pyrrolidones(for example 2-pyrrolidone, 2P), alcohols and alkanols (ethanol, ordecanol), glycols (for example propylene glycol, PG, a common excipientin topically applied dosage forms), surfactants (also common in dosageforms) and terpenes. Other enhancers include oleic acid, oleyl alcohol,ethoxydiglycol, laurocapram, alkanecarboxylic acids, dimethylsulfoxide,polar lipids, or N-methyl-2-pyrrolidone.

In alternative embodiments, the topically active pharmaceutical orcosmetic composition may be optionally combined with other ingredientssuch as moisturizers, cosmetic adjuvants, anti-oxidants, chelatingagents, surfactants, foaming agents, conditioners, humectants, wettingagents, emulsifying agents, fragrances, viscosifiers, buffering agents,preservatives, sunscreens and the like. In another embodiment, apermeation or penetration enhancer is included in the composition and iseffective in improving the percutaneous penetration of the activeingredient into and through the stratum corneum with respect to acomposition lacking the permeation enhancer. Various permeationenhancers, including oleic acid, oleyl alcohol, ethoxydiglycol,laurocapram, alkanecarboxylic acids, dimethylsulfoxide, polar lipids, orN-methyl-2-pyrrolidone, are known to those of skill in the art.

In another aspect, the composition may further comprise a hydrotropicagent, which functions to increase disorder in the structure of thestratum corneum, and thus allows increased transport across the stratumcorneum. Various hydrotropic agents such as isopropyl alcohol, propyleneglycol, or sodium xylene sulfonate, are known to those of skill in theart. The compositions of this invention may also contain active amountsof retinoids (i.e., compounds that bind to any members of the family ofretinoid receptors), including, for example, tretinoin, retinol, estersof tretinoin and/or retinol and the like.

The composition of the invention may comprise a preservative from about0.005% to 2.0% by total weight of the composition. The preservative isused to prevent spoilage in the case of an aqueous gel because ofrepeated patient use when it is exposed to contaminants in theenvironment from, for example, exposure to air or the patient's skin,including contact with the fingers used for applying a composition ofthe invention such as a therapeutic gel or cream. Examples ofpreservatives useful in accordance with the invention included but arenot limited to those selected from the group consisting of benzylalcohol, sorbic acid, parabens, imidurea and combinations thereof. Aparticularly preferred preservative is a combination of about 0.5% to2.0% benzyl alcohol and 0.05% to 0.5% sorbic acid.

The composition preferably includes an antioxidant and a chelating agentwhich inhibit the degradation of the compound for use in the inventionin the aqueous gel formulation. Preferred antioxidants for somecompounds are BHT, BHA, alpha-tocopherol and ascorbic acid in thepreferred range of about 0.01% to 5% and BHT in the range of 0.01% to 1%by weight by total weight of the composition. Preferably, the chelatingagent is present in an amount of from 0.01% to 0.5% by weight by totalweight of the composition. Particularly preferred chelating agentsinclude edetate salts (e.g. disodium edetate) and citric acid in theweight range of about 0.01% to 0.20% and more preferably in the range of0.02% to 0.10% by weight by total weight of the composition. Thechelating agent is useful for chelating metal ions in the compositionwhich may be detrimental to the shelf life of the formulation. While BHTand disodium edetate are the particularly preferred antioxidant andchelating agent respectively for some compounds, other suitable andequivalent antioxidants and chelating agents may be substitutedtherefore as would be known to those skilled in the art.

Additional components may include, but should not be limited to thoseincluding water, oil (eg., olive oil/PEG7), biovera oil, wax (eg.,jojoba wax), squalene, myristate (eg., isopropyl myristate),triglycerides (eg., caprylic triglyceride), Solulan 98, cocoa butter,shea butter, alcohol (eg., behenyl alcohol), stearate (eg.,glycerol-monostearate), chelating agents (eg., EDTA), propylene glycol,SEPIGEL (Seppic, Inc., Fairfield, N.J.), silicone and siliconederivatives (eg., dimethicone, cyclomethicone), vitamins (eg., vitaminE), among others.

Buccal Administration

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for buccal administration. Suchformulations may, for example, be in the form of tablets or lozengesmade using conventional methods, and may, for example, 0.1 to 20% (w/w)active ingredient, the balance comprising an orally dissolvable ordegradable composition and, optionally, one or more of the additionalingredients described herein. Alternately, formulations suitable forbuccal administration may comprise a powder or an aerosolized oratomized solution or suspension comprising the active ingredient. Suchpowdered, aerosolized, or aerosolized formulations, when dispersed,preferably have an average particle or droplet size in the range fromabout 0.1 to about 200 nanometers, and may further comprise one or moreof the additional ingredients described herein.

Rectal Administration

A pharmaceutical composition of the invention may be prepared, packaged,or sold in a formulation suitable for rectal administration. Such acomposition may be in the form of, for example, a suppository, aretention enema preparation, and a solution for rectal or colonicirrigation.

Suppository formulations may be made by combining the active ingredientwith a non-irritating pharmaceutically acceptable excipient which issolid at ordinary room temperature (i.e., about 20° C.) and which isliquid at the rectal temperature of the subject (i.e., about 37° C. in ahealthy human). Suitable pharmaceutically acceptable excipients include,but are not limited to, cocoa butter, polyethylene glycols, and variousglycerides. Suppository formulations may further comprise variousadditional ingredients including, but not limited to, antioxidants, andpreservatives.

Retention enema preparations or solutions for rectal or colonicirrigation may be made by combining the active ingredient with apharmaceutically acceptable liquid carrier. As is well known in the art,enema preparations may be administered using, and may be packagedwithin, a delivery device adapted to the rectal anatomy of the subject.Enema preparations may further comprise various additional ingredientsincluding, but not limited to, antioxidants, and preservatives.

Additional Administration Forms

Additional dosage forms of this invention include dosage forms asdescribed in U.S. Pat. Nos. 6,340,475; 6,488,962; 6,451,808; 5,972,389;5,582,837 and 5,007,790. Additional dosage forms of this invention alsoinclude dosage forms as described in U.S. Patent Applications Nos.20030147952, 20030104062, 20030104053, 20030044466, 20030039688, and20020051820. Additional dosage forms of this invention also includedosage forms as described in PCT Applications Nos. WO 03/35041, WO03/35040, WO 03/35029, WO 03/35177, WO 03/35039, WO 02/96404, WO02/32416, WO 01/97783, WO 01/56544, WO 01/32217, WO 98/55107, WO98/11879, WO 97/47285, WO 93/18755, and WO 90/11757.

Kits of the Invention

The invention also includes a kit comprising a compound useful withinthe methods of the invention and an instructional material thatdescribes, for instance, administering the compound to a subject as aprophylactic or therapeutic treatment or a non-treatment use asdescribed elsewhere herein. In an embodiment, the kit further comprisesa (preferably sterile) pharmaceutically acceptable carrier suitable fordissolving or suspending the therapeutic composition, comprising thecompound useful within the methods of the invention, for instance, priorto administering the molecule to a subject. Optionally, the kitcomprises an applicator for administering the compound.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents were considered to be within the scope of thisinvention and covered by the claims appended hereto. For example, itshould be understood, that modifications in reaction conditions,including but not limited to reaction times, reaction size/volume, andexperimental reagents, such as solvents, catalysts, pressures,atmospheric conditions, e.g., nitrogen atmosphere, andreducing/oxidizing agents, with art-recognized alternatives and using nomore than routine experimentation, are within the scope of the presentapplication.

It is to be understood that wherever values and ranges are providedherein, all values and ranges encompassed by these values and ranges,are meant to be encompassed within the scope of the present invention.Moreover, all values that fall within these ranges, as well as the upperor lower limits of a range of values, are also contemplated by thepresent application.

The following examples further illustrate aspects of the presentinvention. However, they are in no way a limitation of the teachings ordisclosure of the present invention as set forth herein.

EXAMPLES

The invention is now described with reference to the following Examples.These Examples are provided for the purpose of illustration only, andthe invention is not limited to these Examples, but rather encompassesall variations that are evident as a result of the teachings providedherein.

The materials and methods employed in the experiments and the results ofthe experiments presented in this Example are now described.

Example 1 Inhibition of Aβ₁₋₄₂ Levels

In order to correlate plasma and tissue levels of Compound (1) withreduction of amyloid protein levels, mice were administered Compound (1)orally at various doses (25 mg/kg/day; 50 mg/kg/day; and 100 mg/kg/day)for a period of 21 days. The Aβ₁₋₄₂ levels in the brain of the mice weredetermined (FIG. 20A), using a sandwich ELISA assay.

Mice and humans were orally dosed with Compound (1) and the plasmalevels of this compound were determined as a function of time afteradministration (FIG. 20B). The plasma concentrations of Compound (1)were determined by LC/MS/MS. The data illustrated in FIGS. 20A and 20Bsuggest that the levels of Compound (1) in human plasma upon oraladministration of this compound are greater than the levels of Compound(1) in mice for which a reduction in amyloid protein was observed.

Example 2 Proof of Mechanism Study

For a proof of mechanism study of Compound (1), patients suffering frommild cognitive impairment (MCI) were treated with Compound (1) for 10days. The patients were dosed with 4×60 mg Compound (1) for 10 daysorally, using Posiphen® powder filled into gelatin capsules.Cerebrospinal fluid (CSF) and plasma were drawn from the subjects over a12 hour period on day 0 and day 11.

Pharmacokinetic analysis was performed for Compound (1) and selectedmetabolites: N¹-norposiphen, N⁸-norposiphen, and N¹,N⁸-norposiphen.Pharmacodynamic analysis was performed for APP protein and other ADassociated biomarkers in CSF and plasma: APP, Aβ₁₋₄₀, Aβ₁₋₄₂, N-APP,AChEI, BChEI, Tau/p-Tau, NGF, BDNF, and inflammatory factors.

The following schedule of PK monitoring was implemented for eachspecies:

(a) in the mouse, oral/gavage or ip/injection: plasma and brain wereanalyzed at 90, 120, and 180 minutes after administration of the drug,on days 1, 10, 14 and 21.(b) in the rat, oral/gavage or ip/injection or infusion pump/sc: plasma,brain and CSF were analyzed at 90 minutes after administration of drug,on days 1, 5 and 10 days of infusion.(c) in the dog, oral/gelatin capsules filled with Posiphen powder:plasma was analyzed 0 to 36 hours after administration of the drug.(d) in humans, plasma and CSF were analyzed 0 to 12 hours, afteradministration of the drug.

The binding of Compound (1) and metabolites to brain proteins wasdetermined to be ˜96% (rat and human brain homogenate).

FIG. 21A illustrates the levels of Compound (1) and selected metabolitesin the rat plasma, brain and CSF samples under steady state conditions.FIG. 21B illustrates the levels of Compound (1) and selected metabolitesin the human plasma, brain and CSF samples under steady stateconditions.

The levels of Compound (1) and selected metabolites in human plasma(FIG. 22A) and human CSF (FIG. 22B) were determined. Interestingly, thehalf-life of Compound (1) in human plasma was determined to be about 5hours, whereas the half-life of Compound (1) in human CFS was determinedto be about 12 hours. This was corroborated by measuring half-life ofPosiphen® in brains of mice.

Example 3 Effect of Administration of Posiphen® on Human Biomarkers

Levels of biomarkers were monitored in humans after 10 days ofadministration of Compound (1). The results are summarized in FIG. 23.

Example 4 Proposed Phase II Study

In the proposed Phase II study, very early stage (prodome) AD patientsare treated for 1 year. The study comprises 4 to 5 arms, each one withabout 100 people. The arms comprise (i) Compound (1) plus Aricept andAricept control, or (ii) Compound (1) and Aricept plus placebo controls.CSF biomarkers are monitored, and imaging studies and cognitive testsare performed. Safety of subjects is monitored, and if study meets atleast any two endpoints, it is then progressed to Phase III.

The disclosures of each and every patent, patent application, andpublication cited herein are hereby incorporated herein by reference intheir entirety.

While the invention has been disclosed with reference to specificembodiments, it is apparent that other embodiments and variations ofthis invention may be devised by others skilled in the art withoutdeparting from the true spirit and scope of the invention. The appendedclaims are intended to be construed to include all such embodiments andequivalent variations.

1. An amount of(3aR)-1,3a,8-trimethyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indol-5-ylphenylcarbamate (Compound (1)) or a salt thereof, wherein administeringsaid amount to a subject results in a peak plasma circulating level ofCompound (1) ranging from about 10 ng/mL to about 160 ng/ml in saidsubject.
 2. The amount of claim 1, wherein said peak plasma circulatinglevel ranges from about 80 ng/mL to about 160 ng/ml in said subject. 3.The amount of claim 1, wherein said peak plasma circulating level isreached within about 6 hours after said administering.
 4. The amount ofclaim 3, wherein said peak plasma circulating level is reached withinabout 3 hours after said administering.
 5. The amount of claim 1,wherein the plasma circulating level of Compound (1) is equal to orgreater than about 20 ng/mL for at least 12 hours after saidadministering.
 6. The amount of claim 5, wherein the plasma circulatinglevel of Compound (1) is equal to or greater than about 20 ng/mL for atleast 9 hours after said administering.
 7. The amount of claim 1,wherein said administering results in a peak plasma concentration ofCompound (2) ranging from about 15% to about 30% of the peak plasmaconcentration of Compound (1) in said subject.
 8. The amount of claim 1,wherein said administering results in a peak plasma concentration ofCompound (3) ranging from about 15% to about 30% of the peak plasmaconcentration of Compound (1) in said subject.
 9. The amount of claim 1,wherein said administering results in a peak plasma concentration ofCompound (4) ranging from about 1% to about 9% of the peak plasmaconcentration of Compound (1) in said subject.
 10. The amount of claim1, wherein said administering results in a steady state plasmaconcentration of Compound (1) of at least about 100 ng/ml in saidsubject.
 11. The amount of claim 1, wherein said administering resultsin a steady state plasma concentration of Compound (2) of at least about10 ng/ml in said subject.
 12. The amount of claim 1, wherein saidadministering results in a steady state plasma concentration of Compound(3) of at least about 10 ng/ml in said subject.
 13. The amount of claim1, wherein said administering results in a steady state plasmaconcentration of Compound (4) of at least about 3 ng/ml in said subject.14. The amount of claim 1, wherein said administering results in a brainlevel of Compound (1) that ranges from about 4 to about 10 times theplasma level of Compound (1) in said subject.
 15. The amount of claim14, wherein the brain level of Compound (2) ranges from about 15% toabout 150% of the brain level of Compound (1) in said subject.
 16. Theamount of claim 14, wherein the brain level of Compound (3) ranges fromabout 15% to about 150% of the brain level of Compound (1) in saidsubject.
 17. The amount of claim 14, wherein the brain level of Compound(4) is lower than the brain level of Compound (2) or Compound (3) insaid subject.
 18. The amount of claim 1, wherein said subject is ahuman.
 19. A method of inhibiting production of a neurotoxic aggregatingprotein in a subject, said method comprising administering to saidsubject a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and an amount of Compound (1) or a salt thereof,wherein administering said composition to said subject results in a peakplasma circulating level of Compound (1) ranging from about 10 ng/ml toabout 160 mg/ml in said subject, whereby production of said neurotoxicaggregating protein in said subject is inhibited.
 20. The method ofclaim 16, wherein said neurotoxic aggregating protein is selected fromthe group consisting of APP, Aβ, SOD, Tau, alpha-synuclein (SNCA), NAC,TSE prion, and HTT.
 21. The method of claim 19, wherein said peak plasmacirculating level ranges from about 80 ng/mL to about 160 ng/ml in saidsubject.
 22. The method of claim 19, wherein said peak plasmacirculating level is reached within about 6 hours after saidadministering.
 23. The method of claim 22, wherein said peak plasmacirculating level is reached within about 3 hours after saidadministering.
 24. The method of claim 19, wherein the plasmacirculating level of Compound (1) is equal to or greater than about 20ng/mL for at least 12 hours after said administering.
 25. The method ofclaim 24, wherein the plasma circulating level of Compound (1) is equalto or greater than about 20 ng/mL for at least 9 hours after saidadministering.
 26. The method of claim 19, wherein said administeringresults in a peak plasma concentration of Compound (2) ranging fromabout 15% to about 30% of the peak plasma concentration of Compound (1)in said subject.
 27. The method of claim 19, wherein said administeringresults in a peak plasma concentration of Compound (3) ranging fromabout 15% to about 30% of the peak plasma concentration of Compound (1)in said subject.
 28. The method of claim 19, wherein said administeringresults in a peak plasma concentration of Compound (4) ranging fromabout 1% to about 9% of the peak plasma concentration of Compound a) insaid subject.
 29. The method of claim 19, wherein said administeringresults in a steady state plasma concentration of Compound (1) of atleast about 100 ng/ml in said subject.
 30. The method of claim 19,wherein said administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in saidsubject.
 31. The method of claim 19, wherein said administering resultsin a steady state plasma concentration of Compound (3) of at least about10 ng/ml in said subject.
 32. The method of claim 19, wherein saidadministering results in a steady state plasma concentration of Compound(4) of at least about 3 ng/ml in said subject.
 33. The method of claim19, wherein said administering results in a brain level of Compound (1)that ranges from about 4 to about 10 times the plasma level of Compound(1) in said subject.
 34. The method of claim 33, wherein the brain levelof Compound (2) ranges from about 15% to about 150% of the brain levelof Compound (1) in said subject.
 35. The method of claim 33, wherein thebrain level of Compound (3) ranges from about 15% to about 150% of thebrain level of Compound (1) in said subject.
 36. The method of claim 33wherein the brain level of Compound (4) is lower than the brain level ofCompound (2) or Compound (3) in said subject.
 37. The amount of claim19, wherein said subject is a human.
 38. The method of claim 19, whereinsaid administering results in a reduction equal to or greater than about25% in a cerebrospinal fluid marker selected from the group consistingof sAPP α, sAPP β, Tau and pTau in said subject.
 39. The method of claim38, wherein said administering results in a reduction equal to orgreater than about 30% in a cerebrospinal fluid marker selected from thegroup consisting of sAPP α, sAPP β, Tau and pTau in said subject.
 40. Amethod of treating dementia in a subject, said method comprisingadministering to said subject a pharmaceutical composition comprising apharmaceutically acceptable carrier and an amount of Compound (1) or asalt thereof, wherein administering said composition to said subjectresults in a peak plasma circulating level of Compound (1) ranging fromabout 10 ng/mL to about 160 ng/ml in said subject, whereby said dementiain said subject is treated.
 41. The method of claim 40, wherein saiddementia is Alzheimer's disease.
 42. The method of claim 41, whereinsaid dementia is selected from the group consisting of Parkinson'sdisease, Huntington's disease, Prion's disease, Amyloid LateralSclerosis and a tauopathy.
 43. The method of claim 40, wherein said peakplasma circulating level ranges from about 80 ng/mL to about 160 ng/mlin said subject.
 44. The method of claim 40, wherein said peak plasmacirculating level is reached within about 6 hours after saidadministering.
 45. The method of claim 44, wherein said peak plasmacirculating level is reached within about 3 hours after saidadministering.
 46. The method of claim 40, wherein the plasmacirculating level of Compound (1) is equal to or greater than about 20ng/mL for at least 12 hours after said administering.
 47. The method ofclaim 46, wherein the plasma circulating level of Compound (1) is equalto or greater than about 20 ng/mL for at least 9 hours after saidadministering.
 48. The method of claim 40, wherein said administeringresults in a peak plasma concentration of Compound (2) ranging fromabout 15% to about 30% of the peak plasma concentration of Compound (1)in said subject.
 49. The method of claim 40, wherein said administeringresults in a peak plasma concentration of Compound (3) ranging fromabout 15% to about 30% of the peak plasma concentration of Compound (1)in said subject.
 50. The method of claim 40, wherein said administeringresults in a peak plasma concentration of Compound (4) ranging fromabout 1% to about 9% of the peak plasma concentration of Compound (1) insaid subject.
 51. The method of claim 40, wherein said administeringresults in a steady state plasma concentration of Compound (1) of atleast about 100 ng/ml in said subject.
 52. The method of claim 40,wherein said administering results in a steady state plasmaconcentration of Compound (2) of at least about 10 ng/ml in saidsubject.
 53. The method of claim 40, wherein said administering resultsin a steady state plasma concentration of Compound (3) of at least about10 ng/ml in said subject.
 54. The method of claim 40, wherein saidadministering results in a steady state plasma concentration of Compound(4) of at least about 3 ng/ml in said subject.
 55. The method of claim40, wherein said administering results in a brain level of Compound (1)that ranges from about 4 to about 10 times the plasma level of Compound(1) in said subject.
 56. The method of claim 55, wherein the brain levelof Compound (2) ranges from about 15% to about 150% of the brain levelof Compound (1) in said subject.
 57. The method of claim 55, wherein thebrain level of Compound (3) ranges from about 15% to about 150% of thebrain level of Compound (1) in said subject.
 58. The method of claim 55wherein the brain level of Compound (4) is lower than the brain level ofCompound (2) or Compound (3) in said subject.
 59. The amount of claim40, wherein said subject is a human.
 60. The method of claim 40, whereinsaid administering results in a reduction equal to or greater than about25% in a cerebrospinal fluid marker selected from the group consistingof sAPP α, sAPP β, Tau and pTau in said subject.
 61. The method of claim60, wherein said administering results in a reduction equal to orgreater than about 30% in a cerebrospinal fluid marker selected from thegroup consisting of sAPP α, sAPP β, Tau and pTau in said subject.